Microvascular endothelial cell heterogeneity in the central nervous system

中枢神经系统微血管内皮细胞异质性

基本信息

批准号：
7177092
负责人：
Joel S Pachter
金额：
$ 19.11万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-01-22 至 2008-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7177092
关键词：
Address Affect Area Attention Behavior Biochemistry Biological Blood Blood - brain barrier anatomy Blood Vessels Blood capillaries Brain Capillary Endothelial Cell Cerebral cortex Cerebrum Classification Condition Coupling DNA DNA Microarray Chip DNA Microarray format Data Depth Development Disease Drug Formulations Endothelial Cells Evaluation Exploratory/Developmental Grant Gene Expression Gene Expression Profiling Genes Goals Grant Health Heterogeneity Homeostasis In Situ Inflammatory Investigation Lead Light Location Microarray Analysis Modeling Molecular Molecular Profiling Nature Neuraxis Neurons Parietal Lobe Pattern Peripheral Permeability Physiological Population Process RNA Rate Reporting Reproducibility Research Risk Sampling Source Spinal Cord Staging Support Contracts System Techniques Testing Tissues Transcript Trees United States National Institutes of Health arteriole base capillary design hemodynamics laser capture microdissection neurovascular unit novel tissue culture tool vascular bed venule

项目摘要

DESCRIPTION (provided by applicant): The issue of microvascular endothelial heterogeneity greatly impacts vascular function in both health and disease, and potentially severely complicates the use of tissue culture-based models to study endothelial behavior. Yet, despite this topic having been explored in considerable depth in peripheral vascular beds, surprisingly little attention has yet been focused on the microvascular axis of central nervous system (CNS). Unrelated reports describing particular genes have, nonetheless, provocatively suggested that a diversity in endothelial gene expression may exist at the segmental level (i.e., between arterioles, capillaries and venules) as well as the regional level of the CNS, and approximate or surpass that found in the periphery. Given the preeminence of the microvascular network in forming both the blood-brain barrier and blood- spinal cord barrier, and the significant microvascular involvement in inflammatory, infectious, degenerative and traumatic conditions of the CNS, it is critical that there be a systematic and detailed evaluation of this clinically important issue. It is of further importance that such analysis be performed on microvascular tissue in situ," as endothelial cell gene expression is exquisitely prone to environmental modulation. Accordingly, we propose the following Specific Aims: 1) To validate and optimize the approach of coupling laser capture microdissection (LCM) of brain microvascular endothelial cells, with global gene expression profiling by DMA microarray analysis. This will include demonstrating feasibility, by maximizing the percentage of transcripts detected (P call rate) in capillaries only, and establishing reproducibility, by identifying both technical and biological sources of variance in microarray data; and 2) To use the LCM/microarray approach to compare the global gene expression profiles of endothelial cells from capillaries, venules and arterioles. The R21 format chosen is specifically designed for projects that are exploratory and may involve considerable risk, but lead to the development of novel techniques that could have major impact on a field of research. In this regard, results obtained here will set the stage for evaluating regional gene expression by the neurovascular unit (i.e., endothelial cells and intimately associated neural cells) along the CNS microvascular tree in both health and disease. In turn, these studies will enable formulation of more precise endothelial models to study the molecular basis of physiological and pathophysiological processes of the cerebral microvasculature.

描述（由申请人提供）：微血管内皮异质性的问题极大地影响了健康和疾病中的血管功能，并可能严重使基于组织培养的模型使用基于组织培养的模型来研究内皮行为。然而，尽管该主题在周围血管床中进行了相当大的深度探索，但令人惊讶的是，尚未关注中枢神经系统（CNS）的微血管轴。尽管如此，描述特定基因的无关报告却具有挑衅性地表明，内皮基因表达的多样性可能存在于节段水平（即，在小动脉，毛细血管和静脉里）以及CNS的区域水平以及在外围发现的近似或超过的区域水平。鉴于微血管网络在形成血脑屏障和血脊髓屏障方面的最大程度，以及中枢神经系统的炎症，传染性，退化性和创伤性条件的显着微血管参与，至关重要的是，对这一临床重要问题有系统性评估至关重要。进一步的重要性是在原位对微血管组织进行此类分析，“由于内皮细胞基因表达表达非常容易针对环境调节。因此，我们提出了以下特定目的：1）验证和优化通过脑膜摄取的脑含量分析的激光捕获微型分析的方法，以全局含量的方式依次依次耦合激光捕获的方法。可行性，通过在毛细血管中最大化转录本（p呼叫率），并确定可重复性，通过识别微阵列数据中的技术和生物学来源，并使用LCM/微阵列的方法来比较来自VINELES的全球基因表达概要。具有探索性，可能涉及很大的风险，但会导致新技术的发展，这些技术可能会对研究领域产生重大影响。在这方面，此处获得的结果将为评估沿着健康和疾病中CNS微血管树的神经血管单元（即内皮细胞和密切相关的神经细胞）评估区域基因表达的阶段。反过来，这些研究将使更精确的内皮模型制定，以研究大脑微脉管系统的生理和病理生理过程的分子基础。